A kind of preparation method and application of polycation-coated adenovirus composition

A technology of polycation and adenovirus, which is applied in the direction of non-active ingredients of polymer compounds, medical preparations of non-active ingredients, and pharmaceutical formulas, can solve the problems of reducing the expression efficiency of the genes carried, and achieve the delivery of gene drugs and the effect Stable, long-lasting effect

Inactive Publication Date: 2017-12-12
云南东森生物科技有限公司
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Problems solved by technology

Using in vivo small animal biofluorescence imaging technology, in immune normal mice, we confirmed that the H1-packaged Adv complex significantly improved the transfection efficiency of adenovirus and prolonged the duration of gene expression; especially important: Reuse of H1 / Adv in vivo does not reduce efficiency of gene expression

Method used

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  • A kind of preparation method and application of polycation-coated adenovirus composition
  • A kind of preparation method and application of polycation-coated adenovirus composition
  • A kind of preparation method and application of polycation-coated adenovirus composition

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Experimental program
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Effect test

Embodiment 1H1

[0025] The preparation method of embodiment 1H1 / Adv composition

[0026] H1 (4-10nmol) and Adv5-Luc (1×10 6 , pfu) was dissolved in 5% glucose solution, then 12000 g of low-temperature high-speed centrifugation, and the H1 / Adv5-Luc composition of the precipitated H1 wrapped Adv5-Luc had a diameter of about 100 nanometers and a surface charge close to +30mV ( figure 2 ).

Embodiment 2H

[0027] The intratumoral gene transduction efficiency of embodiment 2H1 / rAdv5-Luc composition

[0028] 1 Establishment of animal models of melanoma

[0029] Mouse melanoma B16-F1 cells (2×10 5 cells / mouse, one-time injection) were planted subcutaneously in immunocompetent mice C57BL / 6. After 7 days, the injection site can be seen to have a diameter of about 5×5mm 2 The bulge is confirmed as a tumor.

[0030] 2. Intratumoral injection: H1 / rAdv5-Luc nano composition (Adv 1.5×10 9 pfu, 6000nmol of amino-group of H1) and rAdv5-Luc (rAdv5-Luc) and the second injection was performed 10 days after the first injection. At 1, 3, 7, 11, and 11 days after the first injection, the expression intensity of luciferase was detected by living small animal biological image technology.

[0031] 3. Tail vein injection: Mice were injected with the H1 / rAdv5-Luc nanocomposition (Adv1.5×10 9 pfu, 6000nmol of amino-group of H1) and rAdv5-Luc (rAdv5-Luc). 1 and 14 days after injection, the ...

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Abstract

The invention provides a composition (H1 / rAdv) gene drug delivery system formed by encapsulating recombinant Adenovirus (rAdv) with a polycationic carrier (H1). The composition (H1 / rAdv) of the present invention can efficiently and targetedly deliver the gene medicine mediated by adenovirus, and is useful for the development of therapeutic, control or preventive genes for genetic diseases, tumors and various refractory diseases. Drug delivery carrier system.

Description

field of invention [0001] The present invention relates to gene drug delivery, in particular to a preparation method for encapsulating an adenovirus (Adv) composition (H1 / Adv) by cyclodextrin-polyethyleneimine-folic acid ternary polymerized polycation carrier (H1). In gene drug delivery and its application in gene therapy. technical background [0002] Gene therapy is a new type of treatment for genetic diseases, tumors and other refractory diseases, which has emerged in the past two decades. It mainly transfers exogenous genetic material to individual cells, and regulates the transcription and expression of genes at the epigenetic level, leading to the expression of gene drugs or the silencing of target genes, so as to achieve therapeutic purposes. With the rapid development of modern biotechnology, the completion of the Human Genome Project, the gradual deepening of human understanding of the nature of diseases, and the gradual elucidation of genes and related diseases, g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K47/40A61K47/46
Inventor 姚宏
Owner 云南东森生物科技有限公司
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